Single sample sequencing service includes

FREE - repeats for failed sequences (Flexi / Premium Run) - use of our Pick-up boxes at various locations Single sample sequencing - universal primers (Flexi / Premium Run) - LGC plasmid DNA standard (simply load and compare 2 μL (100 ng) of plasmid DNA standard to your plasmid preparation) - sample bags for secure sample shipment analytical quality • measur eme nt ac • Automated and standardised ABI 3730 XL cu ra sequencing run with a read length up to 1,100 nt c y

(PHRED20 quality) • re g • Overnight turnaround if samples are delivered u la before 10 am t o r • Stored customer-specific primers are selectable y

t during online ordering for 3 months e s

t i • Return of aliquots of synthesised primers on request. n

g

•

Expert advice and customer support on c h

Tel: +49 (0)30 5304 2230 from 8 am to 6 pm e

Monday to Friday m

i

c

a l

Online ordering system m

e

To place your order please visit our webpage and a

s

u

log onto our online ordering system at r

e

https://shop.lgcgenomics.com m

e

n

t

• Register as a new user •

b

i

• Choose your sequencing service, order labels, o

a

n

manage your data and shipment order a l

y

s

i

s

• Please prepare your samples according to the given •

s

t

a

n

d a r d s

•

f o r e n s i c

t e s t i n

requirements and send your samples in a padded g envelope to us. LGC Ostendstraße 25 • 12459 Berlin • Germany via courier service via post (envelopes only) Tel: +49 (0)30 5304 2230 • Fax: +49 (0)30 5304 2201 LGC Genomics GmbH LGC Genomics GmbH Email: [email protected] Ostendstr. 25 / TGS Haus 8 P.O. Box 940327 Web: www.lgcgenomics.com 12459 Berlin 12443 Berlin Germany Germany 083/3.01-2014-01

For further information please contact us on „ Tel: +49 (0)30 5304 2230 or [email protected] No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, “Ready2 run with flexible solutions and premium quality including photocopying, recording or any retrieval system, without the written permission of the copyright holder. © LGC Limited, 2013. All rights reserved. Please choose between our three services. Template DNA requirements Sample specification The main factors influencing read length are the DNA Please use 1.5 mL tubes (no screw cap) for sample Ready2 Run concentration and the quality of the template. submission. Affix the appropriate label to each tube. Send sufficient volume of template DNA at the • Sequencing of purified PCR products or plasmid DNA • Template DNA must be free of EtOH, EDTA, RNA, salts, concentrations detailed below. • Pre-pipetted primer by customers genomic DNA and proteins • Fixed volume & adjusted DNA concentration • Please use ultrapure water for elution Please note: All sample tubes must be clearly barcode labelled. We cannot otherwise guarantee reliable sample tracking during (see sample specification) • Plasmid DNA should be present in covalently closed sample processing. • Volume: 10 µL DNA + 4 µL primer (see sample and circular form. Volume Volume primer specification) • PCR products need to appear as a single band in an Template DNA Concentration Ready2 Run Flexi • No repetition of failed runs. agarose gel and have to be purified from reaction buffer, primers and nucleotides. Plasmids* 100 ng/ μL • LGC recommends silica membrane-based spin column kits PCR products* DNA + Flexi Run min. 15 Primer for template purification. 200 - 500 bp 10 ng/ μL µL DNA • Sequencing of purifiedPCR products or plasmid DNA 500 - 1,000 bp 20 ng/ μL = 14 µL • Submitted primer in separated tube Custom primer requirements 1,000 - 2,000 bp 40 ng/ μL • > 100 universal primers available or shipment of customer primers • Length of 18 - 25 bases * The sequencing service for Ready2 Run and Flexi Run is optimised • No wobble bases for plasmids up to 10 kb and PCR fragments up to approximately • Up to 15 runs per sample, Volume: min.15 µL (1 run) + 2 kb with normal nucleotide composition. 4 µL for each additional run of template DNA • GC-content of at least 40% • The annealing temperature must be T -3 and should be at • Adjusted DNA concentration (see sample specification) m Sequencing of PCR fragments smaller than 150 bp • Storage of templates, submitted primers, and primers least 52°C is not recommended due to base calling limitations of • To calculate T please use T =4x(G+C)+2x(A+T) synthesised by LGC for 3 months m m the current technology. • 3’- end should be G or C • Repetition of failed runs. • No self-hybridisation (primer dimer, loops) or binding to For an accurate determination of the DNA concentra- Additional service several sites on the template tion of your plasmid please use the LGC plasmid DNA • Primer synthesis • For Ready2 Run: Primer concentration 5 µM ≙ 5 pmol / µL standard which is supplied free of charge. • For Flexi Run: We recommend to ship stock solutions Premium Run (100 µM) dissolved in ultrapure water. Please specify the Sequencing results concentration on the tube. All sequencing data are available for three months • Quality check and DNA quantification via agarose gel • Deprotected from the password-protected download area of the • Complex / difficult templates / large constructs • Without modifications (fluorophore or others) sequencing online ordering system. (e.g Cosmid, BACs) • Free of salts and other contaminants. • Technical support by our experienced scientific team We provide the chromatograms (.ab1 and .scf files) • > 100 universal primers available Plasmid preparation (Premium Run only) and the text sequence (.txt file) extracted by the • Storage of templates, submitted primers and primers LGC software. • Pour 200 - 250 µL of 1.5 % LB-medium agar, together with synthesised by LGC for 3 months the appropriate antibiotic, in a tube. Avoid overfilling and Additionally, the secure download area provides free • Repetition of failed runs. bubbles in the agar. programmes for sequence data visualisation. Additional services • Inoculate single colonies using a sterile toothpick; stab the • Primer design and synthesis toothpick into the agar an incubate at 37°C overnight. • PCR clean-up / plasmid preparation • We do not recommend the transfer of small amounts of a liquid bacterial culture to the tube filled with agar. For further information please contact us on For further information please contact us on Tel: +49 (0)30 5304 2230 or [email protected] Tel: +49 (0)30 5304 2230 or [email protected] Single sample sequencing service includes

FREE - repeats for failed sequences (Flexi / Premium Run) - use of our Pick-up boxes at various locations Single sample sequencing - universal primers (Flexi / Premium Run) - LGC plasmid DNA standard (simply load and compare 2 μL (100 ng) of plasmid DNA standard to your plasmid preparation) - sample bags for secure sample shipment analytical quality • measur eme nt ac • Automated and standardised ABI 3730 XL cu ra sequencing run with a read length up to 1,100 nt c y

(PHRED20 quality) • re g • Overnight turnaround if samples are delivered u la before 10 am t o r • Stored customer-specific primers are selectable y

t during online ordering for 3 months e s

t i • Return of aliquots of synthesised primers on request. n

g

•

Expert advice and customer support on c h

Tel: +49 (0)30 5304 2230 from 8 am to 6 pm e

Monday to Friday m

i

c

a l

Online ordering system m

e

To place your order please visit our webpage and a

s

u

log onto our online ordering system at r

e

https://shop.lgcgenomics.com m

e

n

t

• Register as a new user •

b

i

• Choose your sequencing service, order labels, o

a

n

manage your data and shipment order a l

y

s

i

s

• Please prepare your samples according to the given •

s

t

a

n

d a r d s

•

f o r e n s i c

t e s t i n

requirements and send your samples in a padded g envelope to us. LGC Ostendstraße 25 • 12459 Berlin • Germany via courier service via post (envelopes only) Tel: +49 (0)30 5304 2230 • Fax: +49 (0)30 5304 2201 LGC Genomics GmbH LGC Genomics GmbH Email: [email protected] Ostendstr. 25 / TGS Haus 8 P.O. Box 940327 Web: www.lgcgenomics.com 12459 Berlin 12443 Berlin Germany Germany 083/3.01-2014-01

For further information please contact us on „ Tel: +49 (0)30 5304 2230 or [email protected] No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, “Ready2 run with flexible solutions and premium quality including photocopying, recording or any retrieval system, without the written permission of the copyright holder. © LGC Limited, 2013. All rights reserved. Single sample sequencing service includes

FREE - repeats for failed sequences (Flexi / Premium Run) - use of our Pick-up boxes at various locations Single sample sequencing - universal primers (Flexi / Premium Run) - LGC plasmid DNA standard (simply load and compare 2 μL (100 ng) of plasmid DNA standard to your plasmid preparation) - sample bags for secure sample shipment analytical quality • measur eme nt ac • Automated and standardised ABI 3730 XL cu ra sequencing run with a read length up to 1,100 nt c y

(PHRED20 quality) • re g • Overnight turnaround if samples are delivered u la before 10 am t o r • Stored customer-specific primers are selectable y

t during online ordering for 3 months e s

t i • Return of aliquots of synthesised primers on request. n

g

•

Expert advice and customer support on c h

Tel: +49 (0)30 5304 2230 from 8 am to 6 pm e

Monday to Friday m

i

c

a l

Online ordering system m

e

To place your order please visit our webpage and a

s

u

log onto our online ordering system at r

e

https://shop.lgcgenomics.com m

e

n

t

• Register as a new user •

b

i

• Choose your sequencing service, order labels, o

a

n

manage your data and shipment order a l

y

s

i

s

• Please prepare your samples according to the given •

s

t

a

n

d a r d s

•

f o r e n s i c

t e s t i n

requirements and send your samples in a padded g envelope to us. LGC Ostendstraße 25 • 12459 Berlin • Germany via courier service via post (envelopes only) Tel: +49 (0)30 5304 2230 • Fax: +49 (0)30 5304 2201 LGC Genomics GmbH LGC Genomics GmbH Email: [email protected] Ostendstr. 25 / TGS Haus 8 P.O. Box 940327 Web: www.lgcgenomics.com 12459 Berlin 12443 Berlin Germany Germany 083/3.01-2014-01

For further information please contact us on „ Tel: +49 (0)30 5304 2230 or [email protected] No part of this publication may be reproduced or transmitted in any form or by any means, electronic or mechanical, “Ready2 run with flexible solutions and premium quality including photocopying, recording or any retrieval system, without the written permission of the copyright holder. © LGC Limited, 2013. All rights reserved.